CXCL12 signaling axis in pulmonary arterial heterogeneity, development, and disease

CXCL12 信号轴在肺动脉异质性、发育和疾病中的作用

• 批准号: 10905162
• 负责人: David Brian Frank
• 金额: $ 65.73万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-12 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10905162
• 关键词: Adult; Angiogenesis Pathway; Arteries; Autoimmune Diseases; Automobile Driving; Biological Assay; Blood Vessels; Blood capillaries; CXCR4 gene; Cell surface; Cells; Cellular Assay; Child; Childhood; Chronic Disease; Congenital diaphragmatic hernia; Data; Development; Differentiation and Growth; Disease; Distal; Endothelial Cells; Endothelium; Etiology; Fluorescence-Activated Cell Sorting; Generations; Genes; Genetic; Growth; Heterogeneity; Homeostasis; Hypoxemia; In Vitro; Injury; Label; Lead; Left; Longevity; Lung; Lung diseases; Maintenance; Malignant Neoplasms; Mediator; Methods; Modeling; Molecular; Molecular Target; Mus; Natural regeneration; Ontology; Oxygen; Paper; Pathogenesis; Pathology; Pathway interactions; Pattern; Population; Prevention; Proliferating; Pulmonary Circulation; Pulmonary Hypertension; Pulmonary artery structure; Regulation; Reporter; Role; Series; Signal Transduction; Stromal Cell-Derived Factor 1; Structure of parenchyma of lung; System; Technology; Testing; Tissue Differentiation; Tissues; Trees; Vascular Endothelial Cell; Vascular resistance; Vascularization; Vasodilator Agents; angiogenesis; biobank; cell motility; cellular targeting; disorder prevention; fetal; genetic approach; genetic manipulation; improved; in utero; lung development; lung injury; migration; morphogens; mouse model; nitrofen; novel; novel therapeutics; pharmacologic; programs; pulmonary vascular disorder; receptor; recruit; red fluorescent protein; repaired; response to injury; right ventricular failure; segregation; single cell technology; single-cell RNA sequencing; spatiotemporal; stem cells; targeted treatment; therapeutic target; therapy design; transcriptomics; vascular bed

PROJECT SUMMARY Reengagement of developmental paradigms frequently instructs disease pathogenesis. While critical for growth and differentiation for tissue formation, aberrant expression of these pathways leads to significant pathology such as cancer, autoimmunity, and chronic disease. Similarly, disruption of normal fetal programs in the lung can result in significant pulmonary vascular hypoplasia during development and loss of homeostasis in the adult, suggesting a common role in prevention of disease. Thus, understanding the cellular and molecular mechanisms orchestrating pulmonary vascular development will help identify therapeutic targets for lung disease To investigate the cellular and molecular pathways driving development, we focused on the arterial endothelium, a vascular compartment important in the regulation of vascular resistance and prevention of lung diseases such pulmonary hypertension (PH). We labeled arterial endothelial cells (ECs) using the Cxcl12DsRed fluorescent reporter mouse in which a red fluorescent protein is produced in cells expressing the hallmark arterial gene, Cxcl12. We isolated Cxcl12+ cells and performed single cell RNA sequencing to assess cellular heterogeneity within the pulmonary endothelium. Combined cell annotation, gene ontology analysis, and spatial transcriptomics revealed spatially and functionally distinct novel subpopulations of arterial ECs in the developing lung. In addition, we discovered a CXCL12 morphogen gradient from arteries to capillaries, suggesting a haptotaxis mechanism to pulmonary artery growth. Furthermore, disruption of the morphogen gradient resulted in pulmonary vascular hypoplasia and aberrant branching that we quantified by applying unique methods for the lung. In this proposal, we are assessing an expanded role for CXCL12 in development and disease. Spatial transcriptomics for Cxcl12 and its receptors, Cxcr4 and Ackr3, promote our hypothesis that arterial growth and assembly is governed by a CXCL12 morphogen gradient directing cell migration of capillary progenitor cells to the distal artery. In addition, this pattern of expression is observed into adulthood, and we hypothesize that the pulmonary arterial tree contains spatially and functionally distinct subpopulations conserved throughout the lifespan. We will test these hypotheses using genetic mouse models and single cell technology to uncover mechanisms of arterial heterogeneity and CXCL12-orchestrated pulmonary vascular development. Aim 1. To define the spatiotemporal role of CXCL12 signaling in pulmonary vascular development. Aim 2. To assess CXCL12-dependent EC heterogeneity and signaling in developmental lung disease. Aim 3. To identify and characterize evolutionarily conserved proximal and distal arterial ECs.

项目概要 发育范式的重新参与经常指导疾病的发病机制。虽然对于 组织形成的生长和分化，这些途径的异常表达会导致显着的 病理学，例如癌症、自身免疫和慢性疾病。同样，正常胎儿程序的破坏 肺在发育过程中会导致显着的肺血管发育不全和体内平衡的丧失 成人，表明在预防疾病方面具有共同作用。因此，了解细胞和分子 协调肺血管发育的机制将有助于确定肺部疾病的治疗靶点 为了研究驱动发育的细胞和分子途径，我们重点关注动脉 内皮细胞，是调节血管阻力和预防肺损伤的重要血管室 肺动脉高压（PH）等疾病。我们使用 Cxcl12DsRed 标记动脉内皮细胞 (EC) 荧光报告小鼠，其中表达标志动脉的细胞中产生红色荧光蛋白 基因，Cxcl12。我们分离了 Cxcl12+ 细胞并进行单细胞 RNA 测序来评估细胞 肺内皮内的异质性。结合细胞注释、基因本体分析和空间分析 转录组学揭示了发育中动脉 EC 的空间和功能独特的新亚群 肺。此外，我们发现了从动脉到毛细血管的 CXCL12 形态发生素梯度，这表明 肺动脉生长的趋触机制。此外，形态发生素梯度的破坏导致 在肺血管发育不全和异常分支中，我们通过应用独特的方法来量化 肺。 在本提案中，我们正在评估 CXCL12 在发育和疾病中的扩大作用。空间 Cxcl12 及其受体 Cxcr4 和 Ackr3 的转录组学促进了我们的假设：动脉生长 组装由 CXCL12 形态发生素梯度控制，指导毛细血管的细胞迁移 祖细胞转移到远端动脉。此外，这种表达模式一直持续到成年期，我们 假设肺动脉树包含空间和功能上不同的亚群 在整个生命周期中得到保存。我们将使用遗传小鼠模型和单个 细胞技术揭示动脉异质性和CXCL12协调肺血管的机制 发展。 目标 1. 明确 CXCL12 信号传导在肺血管发育中的时空作用。 目标 2. 评估发育性肺疾病中 CXCL12 依赖性 EC 异质性和信号传导。 目标 3. 识别和表征进化上保守的近端和远端动脉 EC。